Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Oncogenomics
  • Published:

Proteomic analysis of hematopoietic stem cell-like fractions in leukemic disorders

Oncogene volume 22, pages 5720–5728 (2003)Cite this article

Abstract

DNA microarray analysis has been applied to identify molecular markers of human hematological malignancies. However, the relatively low correlation between the abundance of a given mRNA and that of the encoded protein makes it important to characterize the protein profile directly, or ‘proteome,’ of malignant cells in addition to the ‘transcriptome.’ To identify proteins specifically expressed in leukemias, here we isolated AC133+ hematopoietic stem cell-like fractions from the bone marrow of 13 individuals with various leukemic disorders, and compared their protein profiles by two-dimensional electrophoresis. A total of 11 differentially expressed protein spots corresponding to 10 independent proteins were detected, and peptide fingerprinting combined with mass spectrometry of these proteins revealed them to include NuMA (nuclear protein that associates with the mitotic apparatus), heat shock proteins, and redox regulators. The abundance of NuMA in the leukemic blasts was significantly related to the presence of complex karyotype anomalies. Conditional expression of NuMA in a mouse myeloid cell line resulted in the induction of aneuploidy, cell cycle arrest in G2–M phases, and apoptosis. These results demonstrate the potential of proteome analysis with background-matched cell fractions obtained from fresh clinical specimens to provide insight into the mechanism of human leukemogenesis.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3

Similar content being viewed by others

References

  • Adler V, Yin Z, Fuchs SY, Benezra M, Rosario L, Tew KD, Pincus MR, Sardana M, Henderson CJ, Wolf CR, Davis RJ and Ronai Z . (1999). EMBO J., 18, 1321–1334.

  • Alizadeh AA, Eisen MB, Davis RE, Ma C, Lossos IS, Rosenwald A, Boldrick JC, Sabet H, Tran T, Yu X, Powell JI, Yang L, Marti GE, Moore T, Hudson Jr J, Lu L, Lewis DB, Tibshirani R, Sherlock G, Chan WC, Greiner TC, Weisenburger DD, Armitage JO, Warnke R, Levy R, Wilson W, Grever MR, Byrd JC, Botstein D, Brown PO and Staudt LM . (2000). Nature, 403, 503–511.

  • Armstrong SA, Staunton JE, Silverman LB, Pieters R, den Boer ML, Minden MD, Sallan SE, Lander ES, Golub TR and Korsmeyer SJ . (2002). Nat. Genet., 30, 41–47.

  • Compton DA and Cleveland DW . (1993). J. Cell Biol., 120, 947–957.

  • The genome international sequencing consortium. (2001). Nature, 409, 860–921.

  • Davis Jr W, Ronai Z and Tew KD . (2001). J. Pharmacol. Exp. Ther., 296, 1–6.

  • Du Q, Stukenberg PT and Macara IG . (2001). Nat. Cell Biol., 3, 1069–1075.

  • Duggan DJ, Bittner M, Chen Y, Meltzer P and Trent JM . (1999). Nat. Genet., 21, 10–14.

  • Gaglio T, Saredi A and Compton DA . (1997). J. Cell Biol., 131, 693–708.

  • Golub TR, Slonim DK, Tamayo P, Huard C, Gaasenbeek M, Mesirov JP, Coller H, Loh ML, Downing JR, Caligiuri MA, Bloomfield CD and Lander ES . (1999). Science, 286, 531–537.

  • Greenberger JS, Sakakeeny MA, Humphries RK, Eaves CJ and Eckner RJ . (1983). Proc. Natl. Acad. Sci. USA, 80, 2931–2935.

  • Gygi SP, Rochon Y, Franza BR and Aebersold R . (1999). Mol. Cell. Biol., 19, 1720–1730.

  • Jaattela M . (1999). Ann. Med., 31, 261–271.

  • Kaul SC, Duncan EL, Englezou A, Takano S, Reddel RR, Mitsui Y and Wadhwa R . (1998). Oncogene, 17, 907–911.

  • Kaul SC, Wadhwa R, Matsuda Y, Hensler PJ, Pereira-Smith OM, Komatsu Y and Mitsui Y . (1995). FEBS Lett., 361, 269–272.

  • Lian Z, Kluger Y, Greenbaum DS, Tuck D, Gerstein M, Berliner N, Weissman SM and Newburger PE . (2002). Blood, 100, 3209–3220.

  • Lian Z, Wang L, Yamaga S, Bonds W, Beazer-Barclay Y, Kluger Y, Gerstein M, Newburger PE, Berliner N and Weissman SM . (2001). Blood, 98, 513–524.

  • Makishima H, Ishida F, Ito T, Kitano K, Ueno S, Ohmine K, Yamashita Y, Ota J, Ota M, Yamauchi K and Mano H . (2002). Br. J. Haematol., 118, 462–469.

  • Matsumine H, Saito M, Shimoda-Matsubayashi S, Tanaka H, Ishikawa A, Nakagawa-Hattori Y, Yokochi M, Kobayashi T, Igarashi S, Takano H, Sanpei K, Koike R, Mori H, Kondo T, Mizutani Y, Schaffer AA, Yamamura Y, Nakamura S, Kuzuhara S, Tsuji S and Mizuno Y . (1997). Am. J. Hum. Genet., 60, 588–596.

  • Miyazato A, Ueno S, Ohmine K, Ueda M, Yoshida K, Yamashita Y, Kaneko T, Mori M, Kirito K, Toshima M, Nakamura Y, Saito K, Kano Y, Furusawa S, Ozawa K and Mano H . (2001). Blood, 98, 422–427.

  • Ohmine K, Ota J, Ueda M, Ueno S-I, Yoshida K, Yamashita Y, Kirito K, Imagawa S, Nakamura Y, Saito K, Akutsu M, Mitani K, Kano Y, Komatsu N, Ozawa K and Mano H . (2001). Oncogene, 20, 8249–8257.

  • Okuyama T, Maehara Y, Endo K, Baba H, Adachi Y, Kuwano M and Sugimachi K . (1994). Cancer, 74, 1230–1236.

  • Pang XP, Hershman JM and Karsan A . (1997). Oncol. Res., 9, 623–627.

  • Shevchenko A, Wilm M, Vorm O and Mann M . (1996). Anal. Chem., 68, 850–858.

  • Shimoda-Matsubayashi S, Hattori T, Matsumine H, Shinohara A, Yoritaka A, Mori H, Kondo T, Chiba M and Mizuno Y . (1997). Neurology, 49, 1257–1262.

  • Venter JC, Adams MD, Myers EW, Li PW, Mural RJ, Sutton GG, Smith HO, Yandell M, Evans CA, Holt RA, Gocayne JD, Amanatides P, Ballew RM, Huson DH, Wortman JR, Zhang Q, Kodira CD, Zheng XH, Chen L, Skupski M, Subramanian G, Thomas PD, Zhang J, Gabor Miklos GL, Nelson C, Broder S, Clark AG, Nadeau J, McKusick VA, Zinder N, Levine AJ, Roberts RJ, Simon M, Slayman C, Hunkapiller M, Bolanos R, Delcher A, Dew I, Fasulo D, Flanigan M, Florea L, Halpern A, Hannenhalli S, Kravitz S, Levy S, Mobarry C, Reinert K, Remington K, Abu-Threideh J, Beasley E, Biddick K, Bonazzi V, Brandon R, Cargill M, Chandramouliswaran I, Charlab R, Chaturvedi K, Deng Z, Di Francesco V, Dunn P, Eilbeck K, Evangelista C, Gabrielian AE, Gan W, Ge W, Gong F, Gu Z, Guan P, Heiman TJ, Higgins ME, Ji RR, Ke Z, Ketchum KA, Lai Z, Lei Y, Li Z, Li J, Liang Y, Lin X, Lu F, Merkulov GV, Milshina N, Moore HM, Naik AK, Narayan VA, Neelam B, Nusskern D, Rusch DB, Salzberg S, Shao W, Shue B, Sun J, Wang Z, Wang A, Wang X, Wang J, Wei M, Wides R, Xiao C, Yan C, Yao A, Ye J, Zhan M, Zhang W, Zhang H, Zhao Q, Zheng L, Zhong F, Zhong W, Zhu S, Zhao S, Gilbert D, Baumhueter S, Spier G, Carter C, Cravchik A, Woodage T, Ali F, An H, Awe A, Baldwin D, Baden H, Barnstead M, Barrow I, Beeson K, Busam D, Carver A, Center A, Cheng ML, Curry L, Danaher S, Davenport L, Desilets R, Dietz S, Dodson K, Doup L, Ferriera S, Garg N, Gluecksmann A, Hart B, Haynes J, Haynes C, Heiner C, Hladun S, Hostin D, Houck J, Howland T, Ibegwam C, Johnson J, Kalush F, Kline L, Koduru S, Love A, Mann F, May D, McCawley S, McIntosh T, McMullen I, Moy M, Moy L, Murphy B, Nelson K, Pfannkoch C, Pratts E, Puri V, Qureshi H, Reardon M, Rodriguez R, Rogers YH, Romblad D, Ruhfel B, Scott R, Sitter C, Smallwood M, Stewart E, Strong R, Suh E, Thomas R, Tint NN, Tse S, Vech C, Wang G, Wetter J, Williams S, Williams M, Windsor S, Winn-Deen E, Wolfe K, Zaveri J, Zaveri K, Abril JF, Guigo R, Campbell MJ, Sjolander KV, Karlak B, Kejariwal A, Mi H, Lazareva B, Hatton T, Narechania A, Diemer K, Muruganujan A, Guo N, Sato S, Bafna V, Istrail S, Lippert R, Schwartz R, Walenz B, Yooseph S, Allen D, Basu A, Baxendale J, Blick L, Caminha M, Carnes-Stine J, Caulk P, Chiang YH, Coyne M, Dahlke C, Mays A, Dombroski M, Donnelly M, Ely D, Esparham S, Fosler C, Gire H, Glanowski S, Glasser K, Glodek A, Gorokhov M, Graham K, Gropman B, Harris M, Heil J, Henderson S, Hoover J, Jennings D, Jordan C, Jordan J, Kasha J, Kagan L, Kraft C, Levitsky A, Lewis M, Liu X, Lopez J, Ma D, Majoros W, McDaniel J, Murphy S, Newman M, Nguyen T, Nguyen N, Nodell M, Pan S, Peck J, Peterson M, Rowe W, Sanders R, Scott J, Simpson M, Smith T, Sprague A, Stockwell T, Turner R, Venter E, Wang M, Wen M, Wu D, Wu M, Xia A, Zandieh A and Zhu X . (2001). Science, 291, 1304–1351.

  • Volm M, Mattern J and Samsel B . (1992). Cancer, 70, 764–769.

  • Watarai H, Inagaki Y, Kubota N, Fuji K, Nagafune J, Yamaguchi Y and Kadoya T . (2000). Electrophoresis, 21, 460–464.

  • Yamashita Y, Miyazato A, Ohya K, Ikeda U, Shimada K, Miura Y, Ozawa K and Mano H . (1996). Jpn. J. Cancer Res., 87, 1106–1110.

Download references

Acknowledgements

This work was supported in part by a grant for Research on the Human Genome, Tissue Engineering, and Food Biotechnology and a grant for the Second-Term Comprehensive 10-Year Strategy for Cancer Control from the Ministry of Health, Labor, and Welfare of Japan; by the Science Research Promotion Fund of the Promotion and Mutual Aid Corporation for Private Schools of Japan; and by a grant from the Research Foundation for Community Medicine, Japan. JO is a research resident of the Japan Health Sciences Foundation.

Author information

Author notes

  1. Jun Ota and Yoshihiro Yamashita: These two authors contributed equally to this work

Authors and Affiliations

  1. Division of Functional Genomics, Jichi Medical School, 3311-1 Yakushiji, Kawachi-gun, Tochigi, 329-0498, Japan

    Jun Ota, Yoshihiro Yamashita, Hiroyuki Kisanuki, Shin-ichiro Fujiwara, Madoka Ishikawa, Young Lim Choi, Shuichi Ueno, Ruri Ohki, Koji Koinuma, Tomoaki Wada & Hiroyuki Mano

  2. Pharmaceutical Division, Pharmaceutical Research Laboratories, Kirin Brewery Co. Ltd, Takasaki, Gunma, 370-1925, Japan

    Katsuya Okawa

  3. Division of Hematology, Jichi Medical School, Kawachi-gun, Tochigi, 329-0498, Japan

    Shin-ichiro Fujiwara

  4. Division of Cardiology, Jichi Medical School, Kawachi-gun, Tochigi, 329-0498, Japan

    Shuichi Ueno & Ruri Ohki

  5. Department of Surgery, Jichi Medical School, Kawachi-gun, Tochigi, 329-0498, Japan

    Koji Koinuma

  6. Department of Gynecology, Jichi Medical School, Kawachi-gun, Tochigi, 329-0498, Japan

    Tomoaki Wada

  7. Department of Biochemistry, Dartmouth Medical School, Hanover, 03755-3844, NH, USA

    Duane Compton

  8. Product Department, Pharmaceutical Division, R&D Center, Kirin Brewery Co. Ltd, Maebashi, Gunma, 371-0853, Japan

    Toshihiko Kadoya

Authors
  1. Jun Ota
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yoshihiro Yamashita
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Katsuya Okawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hiroyuki Kisanuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Shin-ichiro Fujiwara
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Madoka Ishikawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Young Lim Choi
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Shuichi Ueno
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Ruri Ohki
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Koji Koinuma
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Tomoaki Wada
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Duane Compton
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Toshihiko Kadoya
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Hiroyuki Mano
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hiroyuki Mano.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ota, J., Yamashita, Y., Okawa, K. et al. Proteomic analysis of hematopoietic stem cell-like fractions in leukemic disorders. Oncogene 22, 5720–5728 (2003). https://doi.org/10.1038/sj.onc.1206855

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1206855

Keywords

This article is cited by

Search

Advanced search

Quick links